1887

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Volume 103, Issue 11

Concentrating all helper protein functions on a single entity allows rescue of recombinant measles virus by transfection of just two plasmids No Access

Abstract

The generation of recombinant measles virus (MeV) from manipulated genomes on plasmid DNA is quite a complex and inefficient process. As a member of the order its single-stranded ssRNA genome in negative sense orientation is not infectious, but requires co-availability of the viral RNA-dependent RNA polymerase L, the polymerase co-factor phosphoprotein P, and the nucleocapsid protein N in defined relative amounts to establish infectious centres in transfected cell cultures that release replication-competent recombinant MeV particles. For this so-called rescue, different rescue systems were developed that rely on at least four different components. In this work, we establish a functional MeV rescue system just being composed of two components: the plasmid encoding the (modified) viral genome, and a one-helper-plasmid bundling all helper functions. In contrast to a rescue-system for Newcastle Disease Virus, another paramyxovirus, co-expression of all helper proteins by the same promoter failed. Instead, adaptation of the strength of the respective promoters to drive each helper gene´s expression to the relative expression found in MeV-infected cells or other rescue systems, which indeed adjusted respective mRNA and protein expression, yielded success, albeit not yet to the same efficacy as the four-component system. Thereby, our study paves the way for the development of easier and, after further optimization, more efficient rescue systems to generate recombinant MeV for e.g. the application as a vaccine platform or oncolytic virus, for example.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): helper plasmids , measles virus , recombinant morbillivirus , reverse genetics and virus rescue
Funding
This study was supported by the:
  • Deutsches Zentrum für Infektionsforschung (Award TTU 01.805)
    • Principle Award Recipient: MichaelD. Mühlebach
© 2022 The Authors
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/content/journal/jgv/10.1099/jgv.0.001815
2022-11-28
2024-04-26
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Concentrating all helper protein functions on a single entity allows rescue of recombinant measles virus by transfection of just two plasmids
J. Gen. Virol. 103, 001815 (2022); https://doi.org/10.1099/jgv.0.001815
/content/journal/jgv/10.1099/jgv.0.001815
/content/journal/jgv/10.1099/jgv.0.001815
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